enzyme 2 hace2 receptor  (Sino Biological)

Journal: BMC Infectious Diseases

Article Title: Characterization of private mutations in the spike protein of SARS-CoV-2 correlates with viral prevalence

doi: 10.1186/s12879-025-11414-3

Figure Lengend Snippet: The L455F mutation increases the binding affinity of the spike protein to ACE2. A Fluorescence imaging of WT/EG.5.1/EG.5.1-L455F spike pseudovirus-infected 293/ACE2 cells at 72 h. B Relative Fluorescence Units (RFU) measured in 293/ACE2 cells infected with WT/EG.5.1/EG.5.1-L455F spike pseudoviruses at 72 h. *, P < 0.05, ****, P < 0.0001. C and D Biolayer interferometry was used to analyze the binding of biotinylated hACE2 to EG.5.1 spike protein or EG.5.1-L455F spike protein. The curves represent the results of a global fit of the data using a 1:1 binding model

Article Snippet: The receptor protein Recombinant Human ACE2 Protein (His & AVI Tag), Biotinylated (Sino Biological, Beijing, China) was immobilized at a concentration of 25nM on streptavidin biosensor (SA) (Sartorius, Germany).

Techniques: Mutagenesis, Binding Assay, Fluorescence, Imaging, Infection

Journal: BMC Infectious Diseases

Article Title: Characterization of private mutations in the spike protein of SARS-CoV-2 correlates with viral prevalence

doi: 10.1186/s12879-025-11414-3

Figure Lengend Snippet: Complex models of the spike protein RBD with hACE2. A Structural representation of the complex between EG.5.1 RBD (yellow) and hACE2 (purple), highlighting the positions of L455 (red) and L456 (grey) in the EG.5.1 RBD. B Structural representation of the complex between EG.5.1-L455F RBD (green) and hACE2 (blue), highlighting the positions of F455 (red) and L456 (grey) in the EG.5.1-L455F RBD

Article Snippet: The receptor protein Recombinant Human ACE2 Protein (His & AVI Tag), Biotinylated (Sino Biological, Beijing, China) was immobilized at a concentration of 25nM on streptavidin biosensor (SA) (Sartorius, Germany).

Techniques:

Journal: International journal of translational medicine (Basel, Switzerland)

Article Title: Organic Dust Exposure Enhances SARS-CoV-2 Entry in a PKC α - and ADAM-17-Dependent Manner

doi: 10.3390/ijtm4030032

Figure Lengend Snippet: ODE single and repetitive exposure increases murine lung soluble ACE levels dependent upon ADAM-17 with associated effect on SARS-CoV-2 pseudovirus infectivity. Scatter plots with bars depict mean with SEM of lung ACE2 levels of ( A ) wild-type (WT) mice following single and repetitive (13 times) ODE exposures and ( B ) humanized ACE2 mice treated with or without TAPI- 1, an ADAM-17 inhibitor, prior to single instillation with saline or 12.5% ODE exposure ( n = 6 mice/group). Humanized ACE2 mice were exposed to a single dose or repetitive doses of ODE prior to SARS-CoV-2 pseudovirus (PV) infection with lungs collected 5 days post-infection. ( C ) Scatter plot with mean and SEM depicted viral titer determined by qPCR ( n = 6–8 mice/group). * p < 0.05, ** p < 0.01, **** p < 0.0001; groups compared using Student’s t -test in ( A ) and one-way ANOVA with Tukey’s post hoc test in ( B , C ).

Article Snippet: Eight-week-old wild-type (WT) or transgenic mice expressing the human ACE2 receptor (Jackson Labs, Bar Harbor, ME USA) were used for all animal experiments.

Techniques: Infection, Saline

Journal: International journal of translational medicine (Basel, Switzerland)

Article Title: Organic Dust Exposure Enhances SARS-CoV-2 Entry in a PKC α - and ADAM-17-Dependent Manner

doi: 10.3390/ijtm4030032

Figure Lengend Snippet: Inhibition of PKCα or ADAM-17 along with ODE treatment synergistically increases membrane ACE2 levels enhancing SARS-CoV-2 pseudovirus entry in BEAS-2B cells in vitro. ( A ) Wild-type (WT) and PKCα-deficient (DN) BEAS-2B cells were treated with Gö 6976 (a PKCα inhibitor), TAPI-1 (an ADAM-17 inhibitor), 0.5% ODE, or a combination for 1 h in vitro. The cells were then collected and stained for flow cytometry analysis of membrane ACE2 expression. ( B ) Treated cells were infected for 48 h with SARS-CoV-2 pseudovirus expressing fluorescent dTomato. The cells were then fixed, stained with Hoechst nuclear stain, and analyzed using Operetta CLS. WT cells treated with both ODE and inhibitor had significantly higher infection than single-treated WT groups. ( C ) Representative flow cytometry images showing ACE2 gating. ( D ) Representative immunofluorescence images from Operetta CLS showing pseudovirus-infected cells (20× magnification; scale bar: 100 μm). Data shown are mean ± SEM; n = 9 per group; experiments were repeated 4 times; ** p < 0.01, *** p < 0.001, **** p < 0.0001 (two-way ANOVA with Tukey’s post hoc test).

Article Snippet: Eight-week-old wild-type (WT) or transgenic mice expressing the human ACE2 receptor (Jackson Labs, Bar Harbor, ME USA) were used for all animal experiments.

Techniques: Inhibition, Membrane, In Vitro, Staining, Flow Cytometry, Expressing, Infection, Immunofluorescence

Journal: International journal of translational medicine (Basel, Switzerland)

Article Title: Organic Dust Exposure Enhances SARS-CoV-2 Entry in a PKC α - and ADAM-17-Dependent Manner

doi: 10.3390/ijtm4030032

Figure Lengend Snippet: Proposed mechanism through which agricultural dust exposure could affect SARS-CoV-2 entry in vitro (created with BioRender.com ). Organic dust exposure (ODE) activates Toll-like receptor 2 (TLR2) and MyD88, which then activates protein kinase C alpha (PKCα). Through intermediates, PKCα activates ADAM-17 which cleaves the ACE2 receptor on the cell membrane producing soluble ACE2. PKCα can be inhibited by the addition of Gö 6976 and ADAM-17 can be inhibited by the addition of TAPI-1. If membrane ACE2 is intact, upon SARS-CoV-2 infection, the viral spike protein can bind the receptor and through unknown intermediates diminish IL-8 release in vitro.

Article Snippet: Eight-week-old wild-type (WT) or transgenic mice expressing the human ACE2 receptor (Jackson Labs, Bar Harbor, ME USA) were used for all animal experiments.

Techniques: In Vitro, Membrane, Infection